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FIGURE 3 Parapineal cells show immunoreactivity against substance P and serotonin during ontogeny. Indirect immunofluorescence against substance P (SP) (A?D) and serotonin (5HT) (E?H) in larval (7 and 14 dpf) and adult (1.5?2 year old) Tg(foxd3::GFP) zebrafish. Images correspond to dorsal views of confocal z-stack maximum projections, with anterior to the top and left to the left, showing the fluorescence signal corresponding to SP (magenta in (A,C)) and 5HT (magenta in (E,G)), or the merge fluorescence signals that also include the GFP of the pineal complex (green) and the DAPI/Hoechst nuclear staining providing the left habenula tissue context (blue) (B,D,F,H). Arrows indicate parapineal neurons immunoreactive to SP and 5HT. Abbreviations: PO (pineal organ at the level of the stalk), PP (parapineal). Samples: 7 dpf SP (n = 5), adult SP (n = 5), 14dpf 5HT (n = 5), adult 5HT (n = 5). Scale bars, 10 µm.